Micro-stamping technology will be developed which permits creation of complex spatial patterns of multiple proteins with micrometer resolution. Novel features are the ability to stamp proteins directly, to stamp multiple proteins on a single substrate, and to use glass substrates common in cell culture work. The scope of the work will demonstrate the power of this technology as applied to the study of cellular neuroscience. However, extrapolations from this work should show it to be a powerful tool in other basic and applied medical and biological sciences, including neural prosthetics, spinal cord regrowth, development, clinical testing, and other areas of biotechnology. The first specific is to greatly improve the efficiency of the technology so that a wide range of researchers may use it easily and at low cost. Included are studies of protein transfer and its optimization under changes in pH, concentration, pressure, and stamp dimensions and material composition. The second aim is to use stamped patterns of several proteins -- uniquely achievable with this technology -- to control the development and position of axons versus dendrites in culture, and the positions of glia versus neurons. This will demonstrate that the stamp technology is a novel means of understanding and testing how particular molecules influence attachment and growth of cells. This result will be an important step toward the creation of in vitro neural circuits with which one can investigate properties of the nervous system. The third specific aim is to combine the pattern technology with microelectrode array to technology so that neural circuit electrical activity can be recorded and stimulated at many individual cells over long periods of time. This combined technology will contribute to our understanding of basic neuroscience. It is hoped that the lessons learned in creating defined neural circuits will be applicable to neural prosthetics and spinal cord injury, where it is essential to control glial and neural and to guide regrowth of axons. The technology should be applicable to numerous testing paradigms, including clinical chemistry, where large numbers of replications of variable number of tests needed to identify biological chemicals and their effects on cell cultures.